Clostridial toxin derivatives and methods for treating pain

Abstract

Agents for treating pain, methods for producing the agents and methods for treating pain by administration to a patient of a therapeutically effective amount of the agent. The agent can include a clostridial neurotoxin, or a component or fragment or derivative thereof, attached to a targeting moiety, wherein the targeting moiety is selected from a group consisting of transmission compounds which can be released from neurons upon the transmission of pain signals by the neurons, and compounds substantially similar to the transmission compounds.

1. A modified  botulinum  toxin comprising:
 a) a first polypeptide region comprising;
 i) a substance P targeting moiety that binds to a substance P receptor, wherein the substance P targeting moiety is a substance P peptide, a substance P precursor, a substance P fragment, or a substance P analogue; and 
 ii) a  botulinum  toxin translocation domain or fragment thereof that translocates a  botulinum  toxin proteolytic domain across an endosome membrane; and 
 
 b) a second polypeptide region comprising a  botulinum  toxin proteolytic domain that cleaves a neurosecretory protein; 
 wherein the H C  domain of the modified  botulinum  toxin is removed or modified in order to reduce the targeting of the modified  botulinum  toxin to the neuromuscular junction. 
 
     
     
       2. The modified  botulinum  toxin according to  claim 1 , wherein the  botulinum  toxin proteolytic domain is selected from the group consisting of  botulinum  toxin serotype A proteolytic domain,  botulinum  toxin serotype B proteolytic domain,  botulinum  toxin serotype C1 proteolytic domain,  botulinum  toxin serotype D proteolytic domain,  botulinum  toxin serotype E proteolytic domain,  botulinum  toxin serotype F proteolytic domain and  botulinum  toxin serotype G proteolytic domain. 
     
     
       3. The modified  botulinum  toxin according to  claim 1 , wherein the  botulinum  toxin translocation domain is selected from the group consisting of  botulinum  toxin serotype A translocation domain,  botulinum  toxin serotype B translocation domain,  botulinum  toxin serotype C1 translocation domain,  botulinum  toxin serotype D translocation domain,  botulinum  toxin serotype E translocation domain,  botulinum  toxin serotype F translocation domain and  botulinum  toxin serotype G translocation domain. 
     
     
       4. The modified  botulinum  toxin according to  claim 1 , wherein the neurosecretory protein is selected from the group consisting of synaptosomal associated protein of 25 kD (SNAP-25), vesicle-associated membrane protein (VAMP) and syntaxin. 
     
     
       5. The modified  botulinum  toxin according to  claim 1 , wherein the  botulinum  toxin translocation domain is covalently attached to the Substance P targeting moiety. 
     
     
       6. The modified  botulinum  toxin according to  claim 5 , wherein the  botulinum  toxin translocation domain is covalently attached to the Substance P targeting moiety through one or more spacer components. 
     
     
       7. The modified  botulinum  toxin according to  claim 6 , wherein the covalently attachment of the spacer components to the  botulinum  toxin translocation domain, the Substance P targeting moiety, or both the  botulinum  toxin translocation domain and the Substance P targeting moiety is by chemical conjugation and wherein the Substance P targeting moiety is SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 and SEQ ID NO: 17. 
     
     
       8. The modified  botulinum  toxin according to  claim 6 , wherein the covalently attachment of the spacer components to the  botulinum  toxin translocation domain, the Substance P targeting moiety, or both the  botulinum  toxin translocation domain and the Substance P targeting moiety is by formation of a fusion protein and wherein the Substance P targeting moiety is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 and SEQ ID NO: 18. 
     
     
       9. The modified  botulinum  toxin according to  claim 5 , wherein the covalently attachment of the  botulinum  toxin translocation domain to the Substance P targeting moiety is by chemical conjugation and wherein the Substance P targeting moiety is SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 and SEQ ID NO: 17. 
     
     
       10. The modified  botulinum  toxin according to  claim 5 , wherein the covalently attachment of the  botulinum  toxin translocation domain to the Substance P targeting moiety is by formation of a fusion protein and wherein the Substance P targeting moiety is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 and SEQ ID NO: 18.